Resveratrol attenuates apoptosis of endothelial progenitor cells induced by tert-butyl hydroperoxide / 中国病理生理杂志

ABSTRACT

AIM:To investigate the effect of resveratrol on the apoptosis of endothelial progenitor cells (EPC) induced by tert-butyl hydroperoxide (tBHP) and the underlying mechanisms.METHODS:Total mononuclear cells were isolated from peripheral blood by density gradient centrifugation, and the cells were cultured on fibronectin-coated culture dishes.After 4 d of culture, attached cells were divided into control group, tBHP group, and resveratrol plus tBHP group (pretreated with resveratrol for 24 h and then cultured with 100 μmol/L tBHP for 6 h).The proliferation and migration abilities of the EPC were assessed by MTT assay, BrdU incorporation assay and modified Boyden chamber assay.The proportion of apoptotic EPC was determined by flow cytometry after staining with fluorescein isothiocyanate-conjugated Annexin V and propodium iodide.The level of reactive oxygen species (ROS) was determined by H2DCF-DA method.Caspase-3 activity was assay using a caspase-3 colorimetric assay kit.The protein levels of cleaved caspase-3, Bcl-2 and Bax were determined by Western blot.RESULTS:Resveratrol decreased EPC apoptosis induced by tBHP in a dose-dependent manner.Moreover, resveratrol increased the proliferation and migration abilities of the EPC.Resveratrol decreased intracellular ROS level, caspase-3 activity and the protein level of cleaved caspase-3.Resveratrol also decreased protein expression of Bax and increased protein expression of Bcl-2.CONCLUSION:Resveratrol attenuates EPC apoptosis induced by oxidative stress, and its mechanisms may be related to protecting the mitochondrial function.